多酚氧化酶催化单宁的氧化聚合反应研究

研究了多酚氧化酶催化单宁的氧化聚合反应情况 ,探索了反应条件及 7种化合物对酶活性的抑制效果 ,对产物进行了红外光谱和紫外光谱测试 ,相对粘度法测定了产物的分子量。结果表明 ,反应的最佳条件是pH为 5 .2 9,温度为 3 0℃ ,在所用抑制剂中 ,硫酸铜的抑制作用最强 ,与单宁酸的IR图相比 ,产物的 -OH吸收带变窄 ,在 12 60cm-1处吸收峰增强 ,而紫外吸收无变化。产物平均分子量为 3 5 41,说明多酚氧化酶能催化单宁的氧化聚合     

Protection against Ischemia-Induced Oxidative Stress Conferred by Vagal Stimulation in the Rat Heart: Involvement of the AMPK-PKC Pathway

Reactive oxygen species (ROS) production is an important mechanism in myocardial ischemia and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is one of major sources of ROS in the heart. Previous studies showed that vagus nerve stimulation (VNS) is beneficial in treating ischemic heart diseases. However, the effect of VNS on ROS production remains elusive. In this study, we investigated the role of VNS onischemia-induced ROS production. Our results demonstrated that VNS alleviated the myocardial injury, attenuated the cardiac dysfunction, reserved the antioxidant enzyme activity and inhibited the formation of ROS as evidenced by the decreased NADPH oxidase (Nox) activity and superoxide fluorescence intensity as well as the expression of p67phox, Rac1 and nitrotyrosine. Furthermore, VNS resulted in the phosphorylation and activation of adenosine monophosphate activated protein kinase (AMPK), which in turn led to an inactivation of Nox by protein kinase C (PKC); however, the phenomena were repressed by the administration of a muscarinic antagonist atropine. Taken together, these data indicate that VNS decreases ROS via AMPK-PKC-Nox pathway; this may have potential importance for the treatment of ischemic heart diseases.     

Amperometric Cholesterol Biosensor Using Layer-by-Layer Adsorption Technique on Polyaniline-coated Polyester Films

An amperometric cholesterol biosensor was fabricated using polyaniline-coated polyester films. Polyaniline was dissolved in chloroform with camphorsulfonic acid, and polystyrene was added to this solution. Using this mixed solution, the coating was placed onto polyester films. Cholesterol oxidase was immobilized onto these films using an electrostatic layer-by-layer adsorption technique. Poly(diallyldimethylammonium chloride) was used as the counter ion source. The level of adsorption was examined and evidence of layer-by-layer adsorption was investigated using a quartz crystal microbalance (QCM). A cholesterol biosensor was fabricated from these films as a working electrode, and it was used to measure the cholesterol concentration.     

新型电致发光材料1，5—萘二胺衍生物的合成和性质研究

合成了一种新型的有机电致发光材料：N,N‘－二苯基－N,N‘－二（1－萘基）－1,5－萘二胺（NPN）,测定了其吸收谱和荧光发射光谱。该材料具有很好的热稳定性,DSC测定其玻璃化温度（Tg）高达127℃,循环伏安法（CV）测定其电离热（Ip）为5.30eV,可望用作有机电致发光空穴传或蓝色发光材料。     

苯嘧磺草胺的生物学特性与使用

原卟啉原氧化酶是叶绿素与亚铁原卟啉合成之前四吡咯生物合成途径中最后一种关键酶,此种酶是许多除草剂的作用靶标;原卟啉原氧化酶抑制除草剂施用后导致植物细胞过氧化及光下组织白化。介绍了原卟啉原氧化酶抑制除草剂的一个重要新品种—苯嘧磺草胺的特性及其使用。     

A New Potent and Selective Monoamine Oxidase-B Inhibitor with Extended Conjugation in a Chalcone Framework: 1-[4-(Morpholin-4-yl)phenyl]-5-phenylpenta-2,4-dien-1-one

The general blueprint for the design of monoamine oxidase-B (MAO-B) inhibitors has been based on two phenyl or heteronuclei linked via a spacer of appropriate length. In this study, 1-[4-(morpholin-4-yl)phenyl]-5-phenylpenta-2,4-dien-1-one (MO10) was prepared by the condensation of 4′-morpholinoacetophenone and cinnamaldehyde in basic alcoholic medium. MO10 was assessed for inhibitory activity against two human MAO isoforms, MAO-A and MAO-B. Interestingly, MO10 showed a remarkable inhibition against MAO-B with an IC₅₀ value of 0.044 μM along with a selectivity index of 366.13. The IC₅₀ value was better than that of lazabemide (IC₅₀ value of 0.063 μM), which was used as a reference. Kinetics studies revealed that MO10 acted as a competitive inhibitor of MAO-B, with a K_i value of 0.0080 μM. The observation of recovery of MAO-B inhibition, compared to reference levels showed MO10 to be a reversible inhibitor. MTT assays showed that MO10 was nontoxic to normal VERO cells with an IC₅₀ value of 195.44 μg/mL. SwissADME predicted that MO10 provided advantageous pharmacokinetics profiles for developing agents acting on the central nervous system, that is, high passive human gastrointestinal absorption and blood–brain barrier permeability. Molecular docking simulations showed that MO10 properly entered the aromatic cage formed by Y435, Y398, and FAD of the active site of MAO-B. On the basis of these results, MO10 can be considered a promising starting compound in development of agents for the treatment of various neurodegenerative disorders.     

Substrate Flexibility of the Flavin-Dependent Dihydropyrrole Oxidases PigB and HapB Involved in Antibiotic Prodigiosin Biosynthesis

In the biosynthesis of the tripyrrolic pigment prodigiosin, PigB is a predicted flavin-dependent oxidase responsible for the formation of 2-methyl-3-amylpyrrole (MAP) from a dihydropyrrole. To prove which dihydropyrrole is the true intermediate, both possibilities, 5-methyl-4-pentyl-3,4-dihydro-2H-pyrrole ( 5 a , resulting from transamination of the aldehyde of 3-acetyloctanal) and 2-methyl-3-pentyl-3,4-dihydro-2H-pyrrole ( 6 , resulting from transamination of the ketone), were synthesised. Only 5 a restored pigment production in a strain of Serratia sp. ATCC 39006 blocked earlier in MAP biosynthesis. PigB is membrane-associated and inactive when its transmembrane domain was deleted, but HapB, its homologue in Hahella chejuensis, lacks the transmembrane domain and is active in solution. Two colourimetric assays for PigB and HapB were developed, and the HapB-catalysed reaction was kinetically characterised. Ten analogues of 5 a were synthesised, varying in the C2 and C3 side chains, and tested as substrates of HapB in vitro and for restoration of pigment production in Serratia ΔpigD in vivo. All lengths of side chain tested at C3 were accepted, but only short side chains at C2 were accepted. The knowledge that 5 a is an intermediate in prodigiosin biosynthesis and the ease of synthesis of analogues of 5 a makes a range of prodigiosin analogues readily available by mutasynthesis.     

Hypoxia-Responsive Class III Peroxidases in Maize Roots: Soluble and Membrane-Bound Isoenzymes

Flooding induces low-oxygen environments (hypoxia or anoxia) that lead to energy disruption and an imbalance of reactive oxygen species (ROS) production and scavenging enzymes in plants. The influence of hypoxia on roots of hydroponically grown maize (Zea mays L.) plants was investigated. Gene expression (RNA Seq and RT-qPCR) and proteome (LC–MS/MS and 2D-PAGE) analyses were used to determine the alterations in soluble and membrane-bound class III peroxidases under hypoxia. Gel-free peroxidase analyses of plasma membrane-bound proteins showed an increased abundance of ZmPrx03, ZmPrx24, ZmPrx81, and ZmPr85 in stressed samples. Furthermore, RT-qPCR analyses of the corresponding peroxidase genes revealed an increased expression. These peroxidases could be separated with 2D-PAGE and identified by mass spectrometry. An increased abundance of ZmPrx03 and ZmPrx85 was determined. Further peroxidases were identified in detergent-insoluble membranes. Co-regulation with a respiratory burst oxidase homolog (Rboh) and key enzymes of the phenylpropanoid pathway indicates a function of the peroxidases in membrane protection, aerenchyma formation, and cell wall remodeling under hypoxia. This hypothesis was supported by the following: (i) an elevated level of hydrogen peroxide and aerenchyma formation; (ii) an increased guaiacol peroxidase activity in membrane fractions of stressed samples, whereas a decrease was observed in soluble fractions; and (iii) alterations in lignified cells, cellulose, and suberin in root cross-sections.     

Ethanol shock enhances the recovery of anthocyanin from lowbush blueberry

Deactivation of polyphenol oxidase (PPO) in natural products is essential for downstream processing of functional molecules used as food or food additives, particularly those served as antioxidants. In the present work, we identified two proteins with PPO activity from lowbush blueberry using ammonium sulphate precipitation and chromatography procedures. Deactivation of these proteins was studied using aqueous solutions of ethanol of different concentrations. The PPO activity was recovered after ethanol removal for the protein samples previously soaked in a low concentration ethanol solution. A complete and unrecoverable deactivation of the proteins was achieved using ethanol with concentration over 70% (v/v), as manifested by the significant changes in circular dichroism (CD) and fluorescence spectroscopy measurements. Based on these findings, we propose a new extraction process for blueberry anthocyanin, in which an ethanol shock, i.e. soaking blueberry fruit in a 70% (v/v) ethanol solution for 1 h, is implemented before subsequent procedures. This new process increases the anthocyanin yield by 55% in comparison to that without the ethanol shock.     

多铜氧化酶在大肠杆菌中的分泌表达

生物胺是存在于发酵食品中的一类有机物,过量摄入会危害人体健康。多铜氧化酶中的某些酶具有降解多种生物胺的活性,在减控发酵食品中的氨(胺)类危害物方面具有良好的应用前景。研究多铜氧化酶的分泌表达,对酶的特性改造和工业化生产与应用具有重要意义。本研究通过在解淀粉芽孢杆菌来源的多铜氧化酶N端融合信号肽PhoA实现了多铜氧化酶在大肠杆菌中的分泌表达,胞外酶活为69.8 U/L。通过优化诱导条件和酶的分泌确定了多铜氧化酶最优发酵条件为诱导温度25℃、IPTG浓度0.05 mmol/L、诱导时菌体OD600=1.0、诱导6 h后添加150 mmol/L甘氨酸;发酵40 h时胞外多铜氧化酶酶活达到238.1 U/L,是优化前的3.4倍。